Difference between revisions of "Part:BBa K584019"
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AFP normally is expressed intracellularly to prevent freezing of the cytoplasm, but in our system, it was essential that AFP was extracellularly anchored at the cell membrane, thereby preventing ice formation outside the cell. Therefore, we merged the AFP gene to OmpA via a flexible linker, thereby ensuring the desired extracellular localization of AFP. | AFP normally is expressed intracellularly to prevent freezing of the cytoplasm, but in our system, it was essential that AFP was extracellularly anchored at the cell membrane, thereby preventing ice formation outside the cell. Therefore, we merged the AFP gene to OmpA via a flexible linker, thereby ensuring the desired extracellular localization of AFP. | ||
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| + | A point mutation at Base Pair number 156 of the HPLC6 sequence was performed by replacing the T in the wild-type coding sequence to C.This mutation was necessary as the sequence CTGCAG in the Wild-type coding sequence corresponded to a Pst1 restriction site. GCT(Wild type) and GCC(1 point mutant) are both valid triplet codons for Alanine. | ||
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<!-- Add more about the biology of this part here | <!-- Add more about the biology of this part here | ||
Revision as of 14:24, 18 September 2011
OmpA_transmembrane_domain + Linker + point_mutant_HPLC6(AFP)
This composite part contains the two sequences OmpA and Point Mutant HPLC6(Antifreeze Protein - AFP) that are linked via a flexible linker.
AFP normally is expressed intracellularly to prevent freezing of the cytoplasm, but in our system, it was essential that AFP was extracellularly anchored at the cell membrane, thereby preventing ice formation outside the cell. Therefore, we merged the AFP gene to OmpA via a flexible linker, thereby ensuring the desired extracellular localization of AFP.
A point mutation at Base Pair number 156 of the HPLC6 sequence was performed by replacing the T in the wild-type coding sequence to C.This mutation was necessary as the sequence CTGCAG in the Wild-type coding sequence corresponded to a Pst1 restriction site. GCT(Wild type) and GCC(1 point mutant) are both valid triplet codons for Alanine.
Sequence and Features
- 10COMPATIBLE WITH RFC[10]
- 12COMPATIBLE WITH RFC[12]
- 21COMPATIBLE WITH RFC[21]
- 23COMPATIBLE WITH RFC[23]
- 25COMPATIBLE WITH RFC[25]
- 1000COMPATIBLE WITH RFC[1000]